Abstract：
Paradoxical Raf activation via Raf dimerization is a major drawback of wild/mutant B-Raf inhibitors. Herein, we report that CB-1 a novel, potent B-Raf/c-Raf dual inhibitor, effective against colon cancer cells, irrespective of their genetic status. High-throughput virtual screening of the ChemBridge library against wild B-Raf (B-RafWT), mutant B-Raf (B-RafV600E), and c-Raf was performed using an automated protocol with the AutoDock-VINA. Caco-2 and HT-29 cells were used. Of the 23,365 compounds screened computationally, CB-1 showed the highest binding energy towards B-RafWT with a ΔGbinding score of - 13.0 kcal/mol. The compound was also predicted to be effective against B-RafV600E and c-Raf molecules with ΔGbinding energies of - 10.6 and - 10.1 kcal/mol, respectively. The compound inhibited B-RafWT, B-RafV600E and c-Raf kinases with IC50 values of 27.13, 51.70, and 40.23 nM, respectively. The GI50 value of CB-1 was 247.9 nM in B-RafWT-expressing Caco-2 cells and 352.4 nM in B-RafV600E-expressing HT-29 cells. Dose-dependent increases in total apoptosis and G1 cell cycle phase arrest was observed in CB-1-treated colon cancer cells. The compound decreased B-Raf expression in both wild and mutant colon cancer cells. CB-1, a novel, potent dual B-Raf/c-Raf inhibitor was effective against colon cancer cells bearing wild-type and mutant variants of B-Raf expression.
摘要：
经由Raf二聚化的矛盾的Raf活化是野生/突变型B-Raf抑制剂的主要缺点。在这里，我们报告说CB-1是一部小说，强效B-Raf/c-Raf双重抑制剂，对结肠癌细胞有效，不管他们的基因状况如何。针对野生B-Raf（B-RafWT）的ChemBridge文库的高通量虚拟筛选，突变体B-Raf(B-RafV600E),和c-Raf使用自动方案与AutoDock-VINA进行。使用Caco-2和HT-29细胞。在通过计算筛选的23,365个化合物中，CB-1显示对B-RafWT的最高结合能,ΔG结合评分为-13.0kcal/mol。还预测该化合物对B-RafV600E和c-Raf分子有效，ΔG结合能为-10.6和-10.1kcal/mol，分别。该化合物抑制B-RafWT,B-RafV600E和c-Raf激酶，IC50值分别为27.13、51.70和40.23nM，分别。CB-1的GI50值在表达B-RafWT的Caco-2细胞中为247.9nM,在表达B-RafV600E的HT-29细胞中为352.4nM。在CB-1处理的结肠癌细胞中观察到总凋亡和G1细胞周期期停滞的剂量依赖性增加。该化合物降低了B-Raf在野生型和突变型结肠癌细胞中的表达。CB-1小说,有效的双重B-Raf/c-Raf抑制剂对携带B-Raf表达的野生型和突变型的结肠癌细胞有效。
